Functional Analysis of Mutations in Exon 9 of NF1 Reveals the Presence of Several Elements Regulating Splicing

Neurofibromatosis type 1 (NF1) is one of the most common human hereditary disorders, predisposing individuals to the development of benign and malignant tumors in the nervous system, as well as other clinical manifestations. NF1 is caused by heterozygous mutations in the NF1 gene and around 25% of the pathogenic changes affect pre-mRNA splicing. Since the molecular mechanisms affected by these mutations are poorly understood, we have analyzed the splicing mutations identified in exon 9 of NF1, which is particularly prone to such changes, to better define the possible splicing regulatory elements. Using a minigene approach, we studied the effect of five splicing mutations in this exon described in patients. These highlighted three regulatory motifs within the exon. An in vivo splicing analysis of an extensive collection of changes generated in the minigene demonstrated that the CG motif at c.910-911 is critical for the recognition of exon 9. We also found that the GC motif at c.945-946 is involved in exon recognition through SRSF2 and that this motif is part of a Composite Exon Splicing Regulatory Element made up of physically overlapping enhancer and silencer elements. Finally, through an in vivo splicing analysis and in vitro binding assays, we demonstrated that the c.1007G>A mutation creates an Exonic Splicing Silencer element that binds the hnRNPA1 protein. The complexity of the splicing regulatory elements present in exon 9 is most likely responsible for the fact that mutations in this region represent 25% of all exonic changes that affect splicing in the NF1 gene.     

Occurrence of grapevine chrome mosaic nepovirus in Austria

A virus recovered by inoculation of sap from Austrian vines with yellow mosaic symptoms was compared with, and found virtually indistinguishable from, an authentic Hungarian isolate of grapevine chrome mosaic nepovirus. This seems to be the first record of the virus in Austria.     

Expected trends and surprises in the Lateglacial and Holocene vegetation history of the Iberian Peninsula and Balearic Islands

Recent, high-resolution palaeoecological records are changing the traditional picture of post-glacial vegetation succession in the Iberian Peninsula. In addition to the influence of Lateglacial and Early Holocene climatic changes, other factors are critical in the course of vegetation development and we observe strong regional differences. The floristic composition, location and structure of glacial tree populations and communities may have been primary causes of vegetation development. Refugial populations in the Baetic cordilleras would have been a source, but not the only one, for the early Lateglacial oak expansions. From Mid to Late Holocene, inertial, resilient, and rapid responses of vegetation to climatic change are described, and regional differences in the response are stressed. The role of fire, pastoralism, agriculture, and other anthropogenic disturbances (such as mining), during the Copper, Bronze, Iberian, and Roman times, is analysed. The implications of ecological transitions in cultural changes, especially when they occur as societal collapses, are discussed.     

Adipose Tissue-Derived Stem Cells Reduce Acute and Chronic Kidney Damage in Mice

Acute and chronic kidney injuries (AKI and CKI) constitute syndromes responsible for a large part of renal failures, and are today still associated with high mortality rates. Given the lack of more effective therapies, there has been intense focus on the use stem cells for organ protective and regenerative effects. Mesenchymal stem cells (MSCs) have shown great potential in the treatment of various diseases of immune character, although there is still debate on its mechanism of action. Thus, for a greater understanding of the role of MSCs, we evaluated the effect of adipose tissue-derived stem cells (AdSCs) in an experimental model of nephrotoxicity induced by folic acid (FA) in FVB mice. AdSC-treated animals displayed kidney functional improvement 24h after therapy, represented by reduced serum urea after FA. These data correlated with cell cycle regulation and immune response modulation via reduced chemokine expression and reduced neutrophil infiltrate. Long-term analyses, 4 weeks after FA, indicated that AdSC treatment reduced kidney fibrosis and chronic inflammation. These were demonstrated by reduced interstitial collagen deposition and tissue chemokine and cytokine expression. Thus, we concluded that AdSC treatment played a protective role in the framework of nephrotoxic injury via modulation of inflammation and cell cycle regulation, resulting in reduced kidney damage and functional improvement, inhibiting organ fibrosis and providing long-term immune regulation.     

CD2v Interacts with Adaptor Protein AP-1 during African Swine Fever Infection

African swine fever virus (ASFV) CD2v protein is believed to be involved in virulence enhancement, viral hemadsorption, and pathogenesis, although the molecular mechanisms of the function of this viral protein are still not fully understood. Here we describe that CD2v localized around viral factories during ASFV infection, suggesting a role in the generation and/or dynamics of these viral structures and hence in disturbing cellular traffic. We show that CD2v targeted the regulatory trans-Golgi network (TGN) protein complex AP-1, a key element in cellular traffic. This interaction was disrupted by brefeldin A even though the location of CD2v around the viral factory remained unchanged. CD2v-AP-1 binding was independent of CD2v glycosylation and occurred on the carboxy-terminal part of CD2v, where a canonical di-Leu motif previously reported to mediate AP-1 binding in eukaryotic cells, was identified. This motif was shown to be functionally interchangeable with the di-Leu motif present in HIV-Nef protein in an AP-1 binding assay. However, we demonstrated that it was not involved either in CD2v cellular distribution or in CD2v-AP-1 binding. Taken together, these findings shed light on CD2v function during ASFV infection by identifying AP-1 as a cellular factor targeted by CD2v and hence elucidate the cellular pathways used by the virus to enhance infectivity.     

Determinants of Land Degradation and Fragmentation in Semiarid Vegetation at Landscape Scale

The objective of this paper was to investigate the sensitivity to degradation of semiarid plant communities in terms of plant cover and fragmentation, quantifying relationships between landscape characteristic (physical, socio-economical and historical) and vegetation degradation. The degradation of vegetation was measured as the degree of coverage of the two dominant vegetation types (i.e. tall arid brush and tall grass steppe), while fragmentation was measured as patch size and isolation. Data were obtained using GIS tools, and analyzed by logistic regression and linear multivariate regression. Results showed denser coverage at more elevated, gradual slopes that were not sea-oriented. Historical elements of the landscape had a significant effect on current natural vegetation. According to the fragmentation patterns, the vegetation is in fairly good condition (medium coverage had the largest patches but dense coverage was less isolated) but attention must be given to preserve vegetation, due to the relationships between fragmentation and human activities. Moreover, the protection plan under way in the area appeared to favour denser vegetation cover, while human activities had a measurable effect on vegetation degradation.